GSM6729452: U2OS, 1hdex, input, rep2; Homo sapiens; RIP-Seq
Harvested cells were lysed with a Bioruptor (Diagenode, UCD-200) and GR-HaloTag was pulled down using Magne HaloTag beads (Promega). Input samples were taken before mixing samples with beads. After denaturing and high salt washes, RNA was liberated from the beads using proteinase K (New England Biolabs). RNA was isolated and purified using TRIzol and the RNeasy MinElute cleanup kit (QIAGEN). Input samples were treated the same way excluding bead wash steps. Libraries were prepared using the KAPA RNA HyperPrep kit with RiboErase. Adapters used were IDT xGen UDI-UMI Adapters.